Method for detecting single-chain DNA base mutation by gold nanometer particle
A gold nanoparticle, single-stranded technology, which is applied in the field of detecting single-stranded DNA base mutations using gold nanoparticles, can solve the problems of complicated operation, time-consuming and laborious, high cost, etc., and achieves simplified detection steps, easy mass preparation, and reduced cost. The effect of testing costs
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Embodiment 1
[0023] The detection method was applied to detect the mutation hotspot at codon 273 of p53 gene, the base G was mutated to the base T (see references: Rangel-Lopez A, Maldonado-Rodriguez R, Salcedo-Vargas M, et al.Low density DNAmicroarray for detection of most frequent TP53 missense point mutations. BMC Biotechnol, 2005, 15(8): 1-13), the specific operation steps are as follows:
[0024] 1) Mix 94mL of ultrapure water, 1mL of 1% chloroauric acid solution and 5mL of 1% trisodium citrate solution at room temperature, immediately put it in a microwave oven, heat it on high heat to make it boil, then adjust to medium heat , continue to heat for 8min to maintain the boiling state, cool to room temperature naturally, then centrifuge, concentrate, add water to redisperse, prepare colloidal gold with a particle size of 15nm, the color is dark red and translucent, the prepared colloidal gold TEM electron microscope picture see figure 1 ,Depend on figure 1 It can be seen that the part...
Embodiment 2
[0031] The detection method was applied to detect the mutation hotspot at codon 248 of the p53 gene, and the base G was mutated to the base A (see references: Rangel-Lopez A, Maldonado-Rodriguez R, Salcedo-Vargas M, et al.Low density DNAmicroarray for detection of most frequent TP53 missense point mutations. BMC Biotechnol, 2005, 15(8): 1-13), the specific operation steps are as follows:
[0032]1) Mix 96 mL of ultrapure water, 1 mL of 1% chloroauric acid solution and 3 mL of 1% trisodium citrate solution at room temperature, immediately put it in a microwave oven, add high heat to boil, and then adjust to medium heat , continue to heat and keep boiling for 8min, cool to room temperature naturally, then centrifuge, concentrate, add water to redisperse, and prepare colloidal gold with a particle size of 25nm, bright red and translucent.
[0033] 2) Take 10 fmol of wild-type single-stranded DNA and add it to 50 μL of colloidal gold. After mixing, stand still at room temperature ...
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